基于三代测序技术分析新疆鲜马奶和酸马奶中乳酸菌菌群结构

Analysis of the Structure of Lactic Acid Bacteria in Xinjiang Fresh Mare Milk and Koumiss Using Three Generation Sequencing Technology

传统酸马奶受制作环境和工艺的影响,其菌群结构有所不同。本文运用Pacbio SMRT测序技术,基于乳酸菌特异性引物对新疆部分地区酸马奶和鲜马奶的乳酸菌菌群进行研究,结果表明:鲜马奶中乳酸菌丰富,以瑞士乳杆菌(35.52%)、唾液链球菌(10.11%)、乳酸乳球菌(9.49%)、嗜热链球菌(5.96%)以及德氏乳杆菌(5.37%)为主要乳酸菌菌种。酸马奶以瑞士乳杆菌(61.77%)、马乳酒样乳杆菌(14.91%)、乳酸乳球菌(9.76%)为主要乳酸菌菌种。鲜马奶中腐生葡萄球菌(4.45%)、金黄色葡萄球菌(2.62%)含量显著(P<0.05)高于酸马奶且具有致病性。此外,一些条件致病的链球菌属菌种在鲜马奶中被检出,具有潜在的安全风险,不建议直接饮用。酸马奶中含量较高的瑞士乳杆菌和桥乳杆菌显著正相关(r=0.582,P<0.01)。鲜马奶和酸马奶具有不同的乳酸菌菌群特征,自然发酵的酸马奶品质参差不齐。本文应用的乳酸菌特异性引物为低丰度乳酸菌菌种的挖掘提供参考,为酸马奶的工业化生产提供理论依据。

The microbial community structure of traditional Koumiss is bound to be different because of the microenvironment and production process of each family.In this paper,using Pacbio SMRT sequencing technology,based on lactic acid bacteria specific primers,the lactic acid bacteria flora of fresh mare milk and Koumiss in Zhaosu County and Akto County,Ili Kazak Region,Xinjiang were explored.The results showed that the alpha diversity of lactic acid bacteria in fresh mare milk was significantly higher(P<0.005)than Koumiss,and the lactic acid bacteria in fresh mare milk were more abundant.Lactobacillus helveticus(35.52%),Streptococcus salivarius(10.11%),Lactococcus lactis(9.49%),Streptococcus thermophilus(5.96%)and Lactobacillus delbrueckii(5.37%)were the main species of lactic acid bacteria in fresh mare milk.The main lactic acid bacteria species in Koumiss were Lactobacillus helveticus(61.77%),Lactobacillus kefiranofaciens(14.91%)and Lactococcus lactis(9.76%).Staphylococcus saprophyticus(4.45%)and Staphylococcus aureus(2.62%)whose content in fresh mare milk were significantly higher than Koumiss(P<0.05)were pathogenic.In addition,other pathogenic bacteria were detected in fresh mare milk,such as Streptococcus mitis,Streptococcus agalactiae,Streptococcus gallolyticus,Streptococcus pasteurianus and Streptococcus lutetiensis,which stated that fresh mare milk should not be drink directly.And compared with fresh mare milk,Lactobacillus helveticus and Lactobacillus pontis with higher content in Koumiss were significantly positively correlated(r=0.582,P<0.01).The results of beta diversity stated that fresh mare milk and Koumiss had different lactic acid bacteria flora characteristics,and the dispersion of Koumiss was higher than fresh mare milk,indicating that the quality of traditional Koumiss was uneven.The lactic acid bacteria specific primers used in this article provide better conditions for mining low-abundance lactic acid bacteria,which provides a more adequate theoretical basis for the industrial production of Koumiss.